Journal article

A novel strategy for the targeted analysis of protein and peptide metabolites

Nicholas A Williamson, Charles Reilly, Chor-Teck Tan, Sri-Harsha Ramarathinam, Alun Jones, Christie L Hunter, Francis R Rooney, Anthony W Purcell

Proteomics | WILEY | Published : 2011

Abstract

In many biological applications such as epitope discovery or drug metabolism studies, the detection of naturally processed exogenous proteins (e.g. vaccines or peptide therapeutics) and their metabolites is frequently complicated by the presence of a complex endogenous mixture of closely related or even identical compounds. We describe a method that incorporates stable isotope labelling of the protein of interest, allowing the selective screening of the intact molecule and all metabolites using a modified precursor ion scan. This method involves monitoring the low-molecular-weight fragment ions produced during MS/MS that distinguish isotopically labelled peptides from related endogenous comp..

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University of Melbourne Researchers

Grants

Awarded by Australian Research Council


Awarded by National Health and Medical Research Council of Australia


Awarded by National Institutes of Health


Awarded by NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES


Awarded by NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE


Funding Acknowledgements

The authors thank Dr. Nadine Dudek and Dr. Andrew Webb for critical reading of the manuscript and Dr. Roberto Cappai and Dr. Renwick Dobson for isotopically labelled protein expression. A. W. P. is a National Health and Medical Research Council of Australia Senior Research Fellow. The financial support of the Australian Research Council (Linkage project LP0669856), National Health and Medical Research Council of Australia (Project grant 508929), and National Institutes of Health (R01 grants GM057428-06 and NS036592-09) is gratefully acknowledged.